Corsetti Giovanni, Chen-Scarabelli Carol, Romano Claudia, Pasini Evasio, Dioguardi Francesco S, Onorati Francesco, Knight Richard, Patel Hemang, Saravolatz Louis, Faggian Giuseppe, Scarabelli Tiziano M
Division of Human Anatomy and Physiopathology, Department of Clinical and Experimental Sciences, University of Brescia, Brescia, Italy.
Center for Heart and Vessel Preclinical Studies, Department of Internal Medicine, St. John Hospital and Medical Center, Wayne State University, Detroit, MI, USA.
Med Sci Monit Basic Res. 2019 Feb 4;25:33-44. doi: 10.12659/MSMBR.913436.
BACKGROUND Although originally described as a survival mechanism, it is unknown whether and to what extent autophagy is implicated in the terminal stages of heart failure. Here, we studied magnitude and evolution of autophagy in patients with intractable heart failure. MATERIAL AND METHODS Myocardial samples were obtained from 22 patients with ischemic cardiomyopathy and idiopathic dilated cardiomyopathy who were undergoing cardiac transplantation. Hearts from 11 patients who died from non-cardiac causes were used as control samples. Autophagy was evaluated by immunostaining with a monoclonal microtubule associated protein light chain 3 (LC3)-II antibody, while the relationship of autophagy with apoptosis and oncosis was assessed by double staining with TUNEL (terminal deoxynucleotidyl transferase - mediated deoxyuridine triphosphate nick end labeling) assay and complement 9 (C9) immunological staining, respectively. In addition, several necroptotic markers, including RIP1 and RIP3 (receptor interacting protein kinase 1 and 3), anti-C3 (cleaved-caspase-3), and anti-NF-κB (nuclear factor kappa-light-chain-enhancer of activated B cells) were assessed by immunohistochemistry. RESULTS Anti-LC3-II staining was detected in 8.7±1.6% of the heart failure patient heart samples and in 1.2±0.3% of control patient heart samples. Vacuole formation started at one nuclear pole, before becoming bipolar and involving the cytosol. Subsequently, the autophagic process extended also to the nuclei, which underwent a progressive vacuolization and disintegration, assuming a peculiar "strawberry like appearance". Myocytes with extensive vacuole formation exhibited nuclear degeneration, which was associated with TUNEL, C3, C9, RIP1, and RIP3 positive staining. Conversely, myocytes with less extensive vacuole formation showed RIP1 and NF-κB positive staining, though not positivity for other cell death markers. CONCLUSIONS Autophagy was extensively detected in end-stage heart failure and its progression, resulted in secondary cell death, with occurrence of oncosis and necroptosis exceeding that of apoptosis. Conversely, activation of the RIP1/NF-κB pathway was associated with cell survival.
背景 尽管自噬最初被描述为一种生存机制，但目前尚不清楚自噬是否以及在何种程度上与心力衰竭的终末期有关。在此，我们研究了顽固性心力衰竭患者自噬的程度和演变。 材料与方法 从22例接受心脏移植的缺血性心肌病和特发性扩张型心肌病患者中获取心肌样本。将11例死于非心脏原因患者的心脏用作对照样本。通过用单克隆微管相关蛋白轻链3（LC3）-II抗体进行免疫染色来评估自噬，同时分别通过TUNEL（末端脱氧核苷酸转移酶介导的脱氧尿苷三磷酸缺口末端标记）检测和补体9（C9）免疫染色来评估自噬与凋亡和胀亡的关系。此外，通过免疫组织化学评估几种坏死性凋亡标志物，包括RIP1和RIP3（受体相互作用蛋白激酶1和3）、抗C3（裂解的半胱天冬酶-3）和抗NF-κB（活化B细胞的核因子κ轻链增强子）。 结果 在心力衰竭患者心脏样本中，8.7±1.6%检测到抗LC3-II染色，而在对照患者心脏样本中为1.2±0.3%。液泡形成始于一个核极，然后变为双极并累及细胞质。随后，自噬过程也扩展到细胞核，细胞核经历渐进性空泡化和崩解，呈现出一种特殊的“草莓样外观”。具有广泛液泡形成的心肌细胞表现出核变性，这与TUNEL、C3、C9、RIP1和RIP3阳性染色相关。相反，液泡形成较少的心肌细胞显示RIP1和NF-κB阳性染色，尽管其他细胞死亡标志物为阴性。 结论 在终末期心力衰竭中广泛检测到自噬，其进展导致继发性细胞死亡，胀亡和坏死性凋亡的发生率超过凋亡。相反，RIP1/NF-κB途径的激活与细胞存活相关。
